Improved Survival of Fulminant Liver Failure by Transplantation of Microencapsulated Cryopreserved Porcine Hepatocytes in Mice
Improved Survival of Fulminant Liver Failure by Transplantation of Microencapsulated Cryopreserved Porcine Hepatocytes in Mice
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DOI：
10.3727/096368909788237168
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发表时间：
2009-01-01
影响因子：
3.3
通讯作者：
Buehler, Leo H.
中科院分区：
文献类型：
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作者：
Mei, Jie;Sgroi, Antonino;Buehler, Leo H.
本研究的目的是建立猪肝细胞分离，并评价分离的肝细胞封装，冷冻保存和移植（Tx）在暴发性肝功能衰竭（FLF）小鼠模型的功能。分离猪肝细胞后，用海藻酸钠-聚-L-赖氨酸-海藻酸钠膜微囊化并冷冻保存。在体外，游离和封装肝细胞的白蛋白产生的酶联免疫吸附试验测定。在体内，将包封的肝细胞移植到具有FLF的不同组的小鼠中，并进行以下实验组：组1，空胶囊的Tx;组2，游离原代猪肝细胞的Tx;组3，新鲜包封的猪肝细胞的Tx;组4，冷冻保存的包封的猪肝细胞的Tx。在体外，新鲜或冷冻保存的包囊化猪肝细胞在1周内表现出持续降低的代谢功能（白蛋白和尿素合成，药物代谢）。在体内，第1组和第2组的存活率相似（分别为18%和25%，p > 0.05）。在第3组和第4组中，新鲜或冷冻保存的包囊化猪肝细胞的Tx显著提高存活率，分别为75%和68%（p < 0.05）。原代猪肝细胞经微囊化和冻存后仍能保持其代谢功能。在患有FLF的小鼠中，包封的异种肝细胞的Tx显著提高了存活率。这些结果表明，猪肝细胞可以成功地分离，封装，冷冻保存，并移植到异种受体肝功能衰竭和维持肝脏代谢功能。
The aim of this study was to establish hepatocyte isolation in pigs, and to evaluate function of isolated hepatocytes after encapsulation, cryopreservation, and transplantation (Tx) in a mouse model of fulminant liver failure (FLF). After isolation, porcine hepatocytes were microencapsulated with alginate-poly-L-Lysine-alginate membranes and cryopreserved. In vitro, albumin production of free and encapsulated hepatocytes were measured by enzyme linked-immunoadsorbent assay. In vivo, encapsulated hepatocytes were transplanted into different groups of mice with FLF and the following experimental groups were performed: group 1, Tx of empty capsules; group 2, Tx of free primary porcine hepatocytes; group 3, Tx of fresh encapsulated porcine hepatocytes; group 4, Tx of cryopreserved encapsulated porcine hepatocytes. In vitro, fresh or cryopreserved encapsulated porcine hepatocytes showed a continuous decreasing metabolic function over 1 week (albumin and urea synthesis, drug catabolism). In vivo, groups 1 and 2 showed similar survival (18% and 25%, respectively, p > 0.05). In groups 3 and 4, Tx of fresh or cryopreserved encapsulated porcine hepatocytes significantly increased survival rate to 75% and 68%, respectively (p < 0.05). Primary porcine hepatocytes maintained metabolic functions after encapsulation and cryopreservation. In mice with FLF, Tx of encapsulated xenogeneic hepatocytes significantly improved survival. These results indicate that porcine hepatocytes can successfully be isolated, encapsulated, stored using cryopreservation, and transplanted into xenogeneic recipients with liver failure and sustain liver metabolic functions.